From Bioblast
Description
Abbreviation: ceS,S
Reference: A Optimum digitonin concentration for pce -SUIT-010
SUIT number: D008_ce1;ce2Rot;ce3S;ce4D;1Dig;1c
O2k-Application: O2
- MitoPedia: SUIT
- SUIT-category: ceS,S
- SUIT protocol pattern: diametral ce1;ce2Rot;ce3S;ce4D;1Dig;1c
SUIT-010 O2 pce D008 is designed for the evaluation of optimum digitonin concentration for permeabilizing cells, a requirement to account for differences between cell types, the concentration of cells, and variability between batches of the natural product digitonin.
Communicated by Huete-Ortega M and Gnaiger E (last update 2019-01-22)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce2Rot | ROX | ce1;ce2Rot
| ||
ce3S | SL(n) | S | CII | ce1;ce2Rot;ce3S
|
ce4D | SP | S | CII | ce1;ce2Rot;ce3S;ce4D
|
1Dig | SP | S | CII | ce1;ce2Rot;ce3S;ce4D;1Dig
|
1c | SP | S | CII | ce1;ce2Rot;ce3S;ce4D;1Dig;1c
|
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- Coupling control
- Pathway control
- Β» Electron transfer pathway
- Β» Fatty acid oxidation pathway control state, F
- Β» NADH electron transfer-pathway state, N
- Β» Succinate pathway control state, S
- Β» NS-pathway control state, NS
- Β» Glycerophosphate pathway control state, Gp
- Β» Complex IV single step, CIV
- Β» Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- Digitonin is a natural product and thus the effective concentration has to be determined by titrations for every batch. The optimum effective digitonin concentrations for complete plasma membrane permeabilization can be determined with this protocol.
Compare SUIT protocols
- SUIT-003 have been designed to study the coupling control state of intact cells
- SUIT-003 O2 ce D028 coupling control protocol with succinate for permeability test
- SUIT-006 protocol is a coupling control protocol for mitochondrial preparations
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
Doerrier 2018 Methods Mol Biol | 2018 | Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, MΓ©szΓ‘ros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782:31-70. https://doi.org/10.1007/978-1-4939-7831-1_3 | Human Mouse Rat Saccharomyces cerevisiae | Heart Skeletal muscle Endothelial;epithelial;mesothelial cell Blood cells HEK Platelet |
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry
Labels:
SUIT-010